Methods for fat reduction

ABSTRACT

Disclosed herein are methods, compounds, and compositions for fat reduction, and in particular fat reduction without significant hair growth and/or additional hair growth.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit of U.S. provisional patentapplication 62/154,926 filed on Apr. 30, 2015, which is hereinincorporated by reference in its entirety and serves as the basis of apriority and/or benefit claim for the present application.

FIELD

The present disclosure relates generally to methods for fat reduction,and in particular to therapeutic fat reduction and cosmetic fatreduction which are not accompanied by substantial hair growth oradditional hair growth at the site of fat reduction.

BACKGROUND

Excess fat in an individual can be undesirable for a number of reasons.In some cases, the excess fat can be aesthetically unpleasing, such asin the case of cellulite, and other externally visible fat deposits suchas, for example, fat deposits in the submental (under the chin),abdominal, waist, and thigh regions. In other cases, excess fat canresult in obesity, which can be associated with, and increase thelikelihood of, a myriad of diseases and conditions such as, for exampletype 2 diabetes, sleep apnea, heart disease, some types of cancer,osteoarthritis, and others. Consequently, there is a need for cosmeticfat reduction and therapeutic fat reduction.

Many methods for fat reduction generally involve exercise and dietcontrol. However methods of fat reduction by administration offat-reducing compounds offer advantages such as simplicity, ease ofimplementation, and an ability to target fat deposits in both systemic(e.g. throughout the whole body of the individual) and localized (e.g.directly to submental fat and/or cellulite deposits) manners. Inparticular, the compound bimatoprost is one compound which has beenshown to reduce body weight (see e.g., US Patent ApplicationPublications 2014/0275272 and US 2014/0308354). However, bimatoprostadministration sometimes results in hair growth, an effect exemplifiedby the marketing of bimatoprost formulations as the product LATISSE®.Such hair growth is not always desirable and may in fact sometimes beundesirable.

Therefore, there is a need for methods of fat reduction which do notresult in concomitant hair growth, such as hair growth at the site atwhich fat reduction is desired. Additionally, there is also a need for amethod or treatment in which fat deposits are reduced substantiallywithout the pain and inflammation associated with fat cell freezing,heating, lysing, or otherwise destroying the fat deposits throughphysical or chemical means.

SUMMARY

Described herein are methods for fat reduction, and in particularmethods for fat reduction which do not result in substantial hair growthor additional hair growth at the site of fat reduction.

In one aspect, disclosed herein is a method of reducing body fat in asubject comprising administering to a subject in need thereof aneffective amount compound of Formula I, a compound of Formula II, or amixture thereof:

In another aspect, an effective amount of the compound of formula I isadministered.

In another aspect, an effective amount of the compound of formula II isadministered.

In another aspect, the compound or mixture of compounds is administeredin an amount effective for reducing fat without causing substantial hairgrowth at the site of fat reduction.

In another aspect, the compound or mixture of compounds is administeredin an amount effective for reducing fat without causing additional hairgrowth at the site of fat reduction.

In another aspect, the compound or mixture of compounds is administeredin an amount of about 0.05 mg/kg to about 5 mg/kg.

In another aspect, the compound or mixture of compounds is administeredin an amount of about 0.3 mg/kg to about 5 mg/kg.

In another aspect, the compound or mixture of compounds is administeredtopically, by injection, transdermally, or orally.

In another aspect, the compound or mixture of compounds is administeredto at least one of the subject's submental region, thighs, abdomen, orwaist.

In another aspect, the compound or mixture of compounds is administeredsystemically.

In another aspect, the compound or mixture of compounds is administeredlocally to a fat deposit.

In another aspect, described herein is a pharmaceutical composition foruse in reducing fat comprising an effective amount compound of FormulaI, a compound of Formula II, or a mixture thereof:

and one or more pharmaceutically acceptable excipients.

In another aspect, the composition comprises an effective amount of thecompound of Formula I.

In another aspect, the composition comprises an effective amount of thecompound of Formula II.

In another aspect, the composition reduces fat without causingsubstantial hair growth at the site of fat reduction.

In another aspect, the composition reduces fat without causingadditional hair growth at the site of fat reduction.

In another aspect, the composition comprises the compound or mixture ofcompounds in an amount of about 0.05 mg/kg to about 5 mg/kg.

In another aspect, the composition comprises the compound or mixture ofcompounds in an amount of about 0.3 mg/kg to about 5 mg/kg.

In another aspect, the composition is suitable to be administeredtopically, by injection, transdermally, or orally.

In another aspect, the composition is suitable to be administered to atleast one of the subject's submental region, thighs, abdomen, or waist.

In another aspect, the composition is suitable to be administeredsystemically.

In another aspect, the composition is suitable to be administeredlocally to a fat deposit.

DESCRIPTION OF THE DRAWINGS

FIG. 1A, FIG. 1B, FIG. 1C, and FIG. 1D show the results of agonistresponse in the triplicate impedance assays (FIGS. 1A, 1B, and 1C) andsignature trace of the top dose (FIG. 1D) with human dermal papillacells in an in vitro cell dielectric spectroscopy assay withbimatoprost.

FIG. 2A, FIG. 2B, FIG. 2C, and FIG. 2D show the results of agonistresponse in the triplicate impedance assays (FIGS. 2A, 2B, and 2C) andsignature trace of the top dose (FIG. 2D) with human dermal papillacells in an in vitro cell dielectric spectroscopy assay with thecompound of Formula II as described herein.

FIG. 3A, FIG. 3B, FIG. 3C, and FIG. 3D show the results of agonistresponse in the triplicate impedance assays (FIGS. 3A, 3B, and 3C) andsignature trace of the top dose (FIG. 3D) with human dermal papillacells in an in vitro cell dielectric spectroscopy assay with thecompound of Formula II as described herein.

FIG. 4A and FIG. 4B show the results of an in vivo mouse hair re-growthmodel comparing the compound of Formula I to bimatoprost. FIG. 4A showsthe day hair re-growth commenced and FIG. 4B shows day of full hairgrowth was achieved. These were measured in a masked fashion evaluatedby digital photograph.

FIG. 5A and FIG. 5B show graphs of the weight change in rats upon oraladministration of the compound of Formula I to rats. FIG. 5A shows thegraph of male rat weight change and FIG. 5B shows the graph of femalerat weight change. After day 183, animals were allowed to recoverwithout drug administration up to 30 days.

FIG. 6 shows the results of a pre-adipocyte cell differentiation assayin which 3T3-L1 mouse pre-adipocytes were induced to differentiate inthe presence of bimatoprost, compounds of formula I and II for 3 days atthe concentrations indicated.

DETAILED DESCRIPTION

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the present disclosure. As used herein, theuse of the singular includes the plural unless specifically statedotherwise. As used herein, “or” means “and/or” unless stated otherwise.Furthermore, use of the term “including” as well as other forms, such as“includes,” and “included,” is not limiting. The section headings usedherein are for organizational purposes only and are not to be construedas limiting the subject matter described.

Unless specific definitions are provided, the nomenclatures utilized inconnection with the laboratory procedures and techniques of analyticalchemistry, synthetic organic and inorganic chemistry described hereinare those known in the art. Standard chemical symbols are usedinterchangeably with the full names represented by such symbols. Thus,for example, the terms “hydrogen” and “H” are understood to haveidentical meaning. Standard techniques can be used for chemicalsyntheses, chemical analyses, and formulation.

In some embodiments, the methods described herein include a method ofreducing body fat in a subject comprising administering to a subject inneed of reducing body fat an effective amount compound of Formula I, acompound of Formula II, or a mixture thereof:

The term “body fat” as used herein refers the loose connective tissueknown as “adipose tissue”. Body fat can be present throughout the bodyof an individual, for example, beneath the skin (subcutaneous fat; e.g.cellulite), around internal organs (visceral fat), in bone marrow(yellow bone marrow), in breast tissue (breast fat), around the waist(waist fat; e.g. “love handles”), under the chin (submental fat), thightissue (thigh fat), and other regions of the body as would beidentifiable to a skilled person (e.g. HIV associated lipodystrophy,steatoblepharon, and others).

The amount of body fat in an individual can be determined and/orestimated by a variety of methods identifiable to a skilled person. Forexample, body fat percentage (mass of body fat divided by body mass) canbe estimated by techniques known to a skilled person such as hydrostatic(underwater) weighing, whole-body air displacement plethysmography,near-infrared interactance, dual energy X-ray absorptiometry, bodyaverage density measurement (in conjunction with use of the Brozek orSiri formulas), bioelectrical impedance analysis, anthropometric methods(e.g. skinfold measurements, ultrasound measurements, and estimationsbased on the subject's body mass index), magnetic resonance imaging,computed tomography, and other methods identifiable to a skilled person.Additionally, though not a direct measurement of body fat amount, anindividual's body mass index (BMI) can also be indicative of the amountof body fat in an individual. Additionally, visual inspection can alsoreveal accumulated body fat such as in cellulite which can also be usedas part of a quantitative measurement of cellulite (see, for example,Smalls et al. J. Cosmet. Sci. 2005, 56, 105-120). Additional methods fordetermining and/or estimating the amount of body fat will beidentifiable to a skilled person.

The term “subject” as used herein refers to human or non-human animal. Asubject is “in need of reducing body fat” if the individual desires, isadvised, or otherwise requires a reduction in body fat either fortherapeutic or for cosmetic reasons.

The term “reducing” as in “reducing body fat” as used herein refers to alowering in the amount, mass, or volume of body fat. Such reduction canbe measured and determined by measuring the amount of fat according toone or more of the methods described herein at an initial time pointprior to the administering of the compounds described herein (e.g.compounds of Formulas I or II) and then measuring the amount of body fatat various time points (e.g. during the period of administering thecompounds described herein as well after the administering has ceased).For example, a subject's body weight can be measured prior to beginninga treatment regimen with the compounds described herein and thenmeasured during and after the treatment regimen. A decrease in bodyweight is indicative of a reduction in body fat. Similarly, skinfoldmeasurements and/or other techniques (e.g. magnetic resonance imagingand/or computerized tomography) can be made or performed along with theweight measurements where a decrease in the parameters measured by thosetechniques (i.e. body fat percentage) is indicative of fat reduction.Additionally, the reduction of fat can be determined qualitatively suchas by photographing the whole body, or portions of the body, at varioustime points before, during, and after a treatment regimen where thereduction in fat can be determined by visual inspection of the images(e.g. by seeing a visible reduction and in the size and/or volume of aparticular fat deposit such as submental fat, waist fat, cellulite, andother forms of body fat amenable to visual inspection).

In some embodiments, the reduction of body fat achieved with thecompounds of the present disclosure can occur without substantial hairgrowth or additional hair growth at the site of fat reduction. The term“hair growth” as used herein refers to an increase in the number ofhairs in a given area of skin, an increase in the thickness of the hairsalready present in a given area of skin, and increase in the length ofhairs already present in a given area of skin, or combinations thereof.In particular, the number of hairs in a given area of skin can becounted by visual inspection, by the use of computerized methods such asTrichoScan (trichoscan.com/pages/english/home.php), by punch biopsy, andby other methods identifiable to a skilled person to determine the totalnumber of hairs in a particular region, the total hair density (e.g.number of hairs per square centimeter), and other measurements of thenumber of hairs in a given area of skin identifiable to a skilled person(see, e.g., U. Blume-Peytavi et al. “Hair Growth Assestment Techniques”in Hair Growth and Disorders, U. Blume-Peytavi et al., Eds. Springer:New York 2008, Chapter 8). Similarly, the thickness of hairs alreadypresent in a given area of skin can also be measured by visualinspection, by the use of computerized methods such as TrichoScan, bypunch biopsy, and by other methods identifiable to a skilled person todetermine the thickness of the hairs (e.g. the thickness in μm of thehairs). Similarly, the length of the hairs already present in a givenarea of skin can also be measured by visual inspection, by the use ofcomputerized methods such as TrichoScan, by punch biopsy or by othermethods identifiable to a skilled person to determine the length of thehairs (e.g. the length in mm of the hairs and/or the linear hair growthrate in mm/day). Furthermore, hair thickness and hair length can also becharacterized as a distribution wherein different fractions of hairs canshow different thicknesses and/or lengths which can be plotted as ahistogram which can be used to determine measurements such as averagehair thickness and/or length and cumulative hair thickness and/or length(see, e.g. R. Hoffmann “TrichoScan: a novel tool for the analysis ofhair growth in vivo” J. Investig. Dermatol. Symp. Proc. 2003, 109-115).

In addition, the amount of hair growth measured, whether measurement ofthe number, thickness, or length of hairs, or a combination thereof, canbe used as part of methods of measurement of hair growth such as theFerriman-Gallwey Index (Ferriman D M, Gallwey J D. Clinical assessmentof body hair growth in women. J Clin Endocrinol 1961:21:1440-1447; seealso hirsutism.com/hirsutism-biology/ferriman-gallwey-score.shtm). Inparticular, the Ferriman-Gallwey Index can be used as a representationof hair growth in a male pattern on a woman shown in four differentdegrees of severity in 19 different body regions: namely the upper lip,chin, sideburns, neck, chest, upper back, lower back, buttocks, upperabdomen, lower abdomen, inguinal area, perianal area, arm, forearm,thigh, leg, foot, toes, and fingers wherein a value of 0 (zero)indicates an absence of terminal hair and wherein a value of 1-4 isassigned to each of the 19 regions depending on the amount of hairpresent (see N. F. Goodman et al. “American Association of ClinicalEndocrinologists Medical Guidelines for Clinical Practice for TheDiagnosis and Treatment of Hyperandrogenic Disorders” Endocrine Practice2001, 7, 102-134).

The term “without substantial hair growth” as used herein in connectionwith the fat reduction achieved by administration of compounds describedherein can, in some embodiments, refer to the substantial lack of hairgrowth in a region of skin at the site of fat reduction which did notpreviously have hair present. Additionally, the term “without additionalhair growth” as used herein in connection with the fat reductionachieved by administration of compounds described herein can, in someembodiments, refer to the substantial lack of additional hair growth ina region of skin at the site of fat reduction where at least some hairis already present. In particular, the lack of additional hair growth isrelative to any normal hair growth which would have occurred in absenceof the administration of the compounds described herein. By way ofexample, if one or more of the compounds described herein areadministered to a region which has a certain number of hairs per unitarea (e.g. as determined by methods such as visual inspection and/orcomputerized methods such as TrichoScan, or by determination of aFerriman-Gallwey Index value of 1 or more), then any fat reduction,measured by fat reduction determination methods described herein, can beconsidered to have occurred without additional hair growth if the siteof administration shows a reduction in fat but substantially no newhairs present relative to the state of the region prior toadministration of the compounds (e.g., there will be no hairs present asseen by methods such as visual inspection and/or computerized methodssuch as TrichoScan, and/or the Ferriman-Gallwey Index value would stillbe considered to be the previous value), or if the number of new hairswould not be substantially greater than the normal number of new hairsthat would have been expected if the one or more compounds had not beenadministered. Thus, in some embodiments, even if additional new hairsare counted in addition to the number of hairs already present at thesite prior to administration, if the site would have been expected tohave a normal increase of about 1% to about 10% in the number of hairs(relative to the original number of hairs) if the one or more compoundshad not been administered, then the fat reduction can be considered tooccur without additional hair growth if the administration of the one ormore compounds also resulted in hair growth that was not more than about1% to about 10% (relative to the original number of hairs present priorto the one or more compounds being administered).

Similarly, by way of another example, fat reduction occurring due to theadministration of one or more of the compounds described herein can beconsidered to occur without additional hair growth if the fat reductionoccurs substantially without a change in the thickness of the hair (e.g.the average thickness, cumulative thickness, and/or distribution of hairthicknesses as determined by the visual and/or computerized methodsdescribed herein or as determined as part of the Ferriman-Gallwey Index)relative to the thickness of the hairs at the site of fat reductionprior to the administration of the one or more compounds. Additionally,the fat reduction can also be considered to occur without additionalhair growth if change in thickness of the hair is not substantiallygreater than normal change in hair thickness that would have beenexpected if the one or more compounds had not been administered. Thus,in some embodiments, even if the thickness of the hairs at the siteprior to administration increases, if the hairs at the site would havebeen expected to have a normal thickness increase of about 1% to about10% (relative to the original thickness of hairs) if the one or morecompounds had not been administered, then the fat reduction can beconsidered to occur without additional hair growth if the administrationof the one or more compounds also resulted in hair thickness increasethat was not more than about 1% to about 10% (relative to the originalthickness of hairs prior to the one or more compounds beingadministered).

Similarly, by way of another example, fat reduction occurring due to theadministration of one or more of the compounds described herein can beconsidered to occur without additional hair growth if the fat reductionoccurs without a substantial change in the length of the hair relativeto the length of the hairs (e.g. the average length, cumulative length,and/or distribution of hair lengths as determined by the visual and/orcomputerized methods described herein or as determined as part of theFerriman-Gallwey Index) at the site of fat reduction prior to theadministration of the one or more compounds. Additionally, the fatreduction can also be considered to occur without additional hair growthif change in length of the hair is not greater than normal change inhair length that would have been expected if the one or more compoundshad not been administered. Thus, in some embodiments, even if the lengthof the hairs at the site prior to administration increases, if the hairsat the site would have been expected to have a normal length increase ofabout 1% to about 10% (relative to the original length of hairs) if theone or more compounds had not been administered, then the fat reductioncan be considered to occur without additional hair growth if theadministration of the one or more compounds also resulted in hairthickness increase that was not more than about 1% to about 10%(relative to the original thickness of hairs prior to the one or morecompounds being administered).

A skilled person will also understand that hair growth can be acombination of increases in number of hairs, thickness of hairs, andlengths of hairs. Thus, fat reduction achieved by administration of thecompounds described herein can occur without additional hair growth ifthere is no substantial increase in one or more parameters such asnumber of hairs, thickness of hairs, or length of hairs, and/or if thechange in the one or more parameters is not more than would normally beexpected, as described herein, if the one or more compounds had not beenadministered.

Furthermore, in some embodiments, the fat reduction occurring due to theadministration of one or more compounds described herein can occur withless hair growth (e.g. number of hairs, thickness of hairs, and/orlength of hairs) than occurs with the administration of other compoundswhich cause fat reduction but can also cause hair growth (e.g.bimatoprost). By way of example, administration of the compoundsdescribed herein can be considered to occur without additional hairgrowth if the fat reduction occurs with hair growth at the site of fatreduction that is between 0 and 15%, 0 and 10%, or 0 and 5% of the hairgrowth seen with the administration of the other compound that resultsin a comparable amount of fat reduction (e.g. results in an amount offat reduction that is does not vary by more than about 0 to 20% relativeto the amount of fat reduction achieved with the other compound).

The term “administering” as used herein refers to introduction of asubstance (e.g. the compounds of Formulas I and II described herein)into a body of a subject and/or application of a substance onto the bodyof a subject by a particular route. Routes of administration would beidentifiable to a skilled person and include, for example, oraladministration, parenteral administration (e.g. subcutaneous injection,intramuscular injection, and intravenous injection), sublingualadministration, buccal administration, rectal administration, ocularadministration, optic administration, inhalation routes (e.g. inhaling amist containing the substance though the mouth or nose), topicaladministration, transdermal administration (e.g. via transdermalpatches), administration via an implant device, and others identifiableto a skilled person.

Administration can be “local” when the compound is administered to aparticular localized region of the body and only that region near thesite of administration is exposed to the compound (e.g. topicalapplication or subcutaneous application to a particular region of thesubject's body.)

Similarly, administration can be “systemic” when the compound isadministered such that the compound is exposed throughout the subject'sbody and may be found in one or more regions distant from the site ofadministration (e.g. orally or intravenously administering the compoundsuch that the compound will be distributed in the blood and throughoutvarious tissues and/or body regions resulting in fat reduction at thosetissues and/or regions).

In some embodiments, the compounds can be administered by administeringpharmaceutical compositions to a subject. The pharmaceuticalcompositions can contain at least one compound described herein in apharmaceutically acceptable carrier thereof and can in some embodimentscontain one or more pharmaceutically acceptable excipients. The phrase“pharmaceutically acceptable” means the carrier, diluent or excipient iscompatible with the other ingredients of the formulation and notdeleterious to the recipient thereof.

Pharmaceutical compositions of the compounds described herein can beused in the form of a solid, a solution, an emulsion, a dispersion, apatch, a micelle, a liposome, and others identifiable to a skilledperson, wherein the resulting composition contains one or more compoundsdescribed herein, as an active ingredient, in admixture with an organicor inorganic carrier or excipient suitable for enteral or parenteralapplications. Compounds described herein can be combined, for example,with non-toxic, pharmaceutically acceptable carriers known to a skilledperson for tablets, pellets, capsules, suppositories, solutions,emulsions, suspensions, and any other form suitable for use identifiableto a skilled person. Exemplary carriers which can be used can includeglucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesiumtrisilicate, talc, corn starch, keratin, colloidal silica, potatostarch, urea, medium chain length triglycerides, dextrans, and othercarriers identifiable to a skilled person suitable for use inmanufacturing preparations, in solid, semisolid, or liquid form. Inaddition, auxiliary agents, stabilizing agents, thickening agents,coloring agents, and perfumes can be used. Compounds described hereincan be included in the pharmaceutical composition in an amountsufficient to produce the desired effect upon the subject's condition(e.g. fat reduction as described herein).

Pharmaceutical compositions containing compounds described herein can bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs, as well asothers identifiable to a skilled person. Compositions intended for oraluse can be prepared according to any method known in the art, and othersidentifiable to a skilled person, for the manufacture of pharmaceuticalcompositions and such compositions can contain one or more agentsselected from the group consisting of a sweetening agent such assucrose, lactose, or saccharin, flavoring agents such as peppermint, oilof wintergreen or cherry, coloring agents and preserving agents in orderto provide pharmaceutically elegant and palatable preparations. Tabletscontaining compounds described herein in admixture with non-toxicpharmaceutically acceptable excipients can also be manufactured by knownmethods. The excipients used can be, for example, (1) inert diluentssuch as calcium carbonate, lactose, calcium phosphate or sodiumphosphate, or others identifiable to a skilled person; (2) granulatingand disintegrating agents such as corn starch, potato starch or alginicacid, or others identifiable to a skilled person; (3) binding agentssuch as gum tragacanth, corn starch, gelatin or acacia, or othersidentifiable to a skilled person, and (4) lubricating agents such asmagnesium stearate, stearic acid or talc, or others identifiable to askilled person. The tablets can be uncoated or they can be coated byknown techniques identifiable to a skilled person to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed.

In some cases, formulations for oral use can be in the form of hardgelatin capsules wherein the compounds described herein are mixed withan inert solid diluent, for example, calcium carbonate, calciumphosphate or kaolin, or others identifiable to a skilled person.Formulations can also be in the form of soft gelatin capsules whereinthe compounds described herein are mixed with water or an oil medium,for example, peanut oil, liquid paraffin or olive oil, or othersidentifiable to a skilled person.

The pharmaceutical compositions can be in the form of a sterileinjectable suspension. This suspension can be formulated according toknown methods identifiable to a skilled person using suitable dispersingor wetting agents and suspending agents. The sterile injectablepreparation can also be a sterile injectable solution or suspension in anon-toxic parenterally acceptable diluent or solvent, for example, as asolution in 1,3-butanediol, or other non-toxic parenterally acceptablediluents or solvents identifiable to a skilled person. Sterile, fixedoils are conventionally employed as a solvent or suspending medium. Forthis purpose, a bland fixed oil can be employed including syntheticmono- or diglycerides, fatty acids (including oleic acid), naturallyoccurring vegetable oils like sesame oil, coconut oil, peanut oil,cottonseed oil, and others identifiable to a skilled person, orsynthetic fatty vehicles like ethyl oleate and others identifiable to askilled person. Buffers, preservatives, antioxidants, and othersidentifiable to a skilled person can be incorporated as required.

Pharmaceutical compositions containing compounds described herein can bein a form suitable for topical use, for example, as oily suspensions, assolutions or suspensions in aqueous liquids or non-aqueous liquids, oras oil-in-water or water-in-oil liquid emulsions. Pharmaceuticalcompositions can be prepared by combining a therapeutically effectiveamount of at least one compound described herein as an active ingredientwith conventional topically acceptable pharmaceutical excipients and bypreparation of unit dosage suitable for topical use. Additionalcompositions that may be suitable for use herewith are described in U.S.Patent Application Publication No. 2014/0155488, titled “Compositionsand methods for stimulating hair growth”, the disclosure of which ishereby incorporated by reference.

The compounds described can also be administered in the form ofsuppositories for rectal administration of the compounds. Thesecompositions can be prepared by mixing the compounds described hereinwith a suitable non-irritating excipient, such as, for example, cocoabutter, synthetic glyceride esters of polyethylene glycols, which aresolid at ordinary temperatures, but liquefy and/or dissolve in therectal cavity to release the compounds.

Since individual subjects can present a wide variation amount of fat tobe reduced and each compound has its unique characteristics, the precisemode of administration and dosage employed for each subject is left tothe discretion of the practitioner.

In some embodiments, when the compounds described herein (e.g. compoundsof Formula I and Formula II) are part of a composition, the compoundsare the only active ingredients which result in fat reduction, and inparticular fat reduction without significant hair growth and/oradditional hair growth as described herein. The term “active ingredient”as used herein refers to a component which is responsible for thebiological effect of fat reduction, and in particular fat reductionwithout significant hair growth and/or additional hair growth asdescribed herein, whereas the other components of the composition (e.g.excipients, carriers, and diluents) are not responsible for thebiological effect of fat reduction, and in particular fat reductionwithout significant hair growth and/or additional hair growth asdescribed herein, even if they have other functions in the compositionwhich are necessary or desired as part of the formulation (such aslubrication, flavoring, pH control, emulsification, and other functionsother than fat reduction and in particular fat reduction withoutsignificant hair growth and/or additional hair growth as describedherein).

The term “effective amount” as used herein refers to an amount of acompound (e.g. the compounds of Formula's I and II) which will exert abeneficial effect when administered to an individual. For example, agiven amount of a compound will be an “effective amount” whenadministration of that amount of the compound results in the reductionof fat in a subject as determined by the measurement and evaluationtechniques described herein. In particular, in some embodiments, theeffective amount not only results in reduction of fat, but also does sowithout significant hair growth and/or additional hair growth asdescribed herein and which can be seen with certain compounds such asbimatoprost.

The effective amount can be administered as described herein. Forexample, the effective amount can be formulated as a pharmaceuticalcomposition and injected subcutaneously into a particular deposit of faton the body to reduce the fat in the particular region. Alternatively,the effective amount can be formulated as part of a composition fortopical administration (e.g. a cream or gel) and be topicallyadministered to a particular fat deposit on the subject's body to reducefat in that region. Alternatively, effective amount can be formulated aspart of an oral formulation (e.g. a capsule) or parenteral formulation(e.g. an injectable formulation) and administered to a subject toachieve fat reduction in various regions of a subject's body.

In some embodiments, an effective amount of only a compound of Formula Ican be administered. In other embodiments, an effective amount of only acompound of Formula II can be administered. In other embodiments, acombination of effective amounts of compounds according to Formulas Iand II can be administered.

The actual amount of the compound to be administered in any given casewill be determined by a physician or other skilled person taking intoaccount the relevant circumstances, such as the amount of fat reduction,the age and weight of the patient, the patient's general physicalcondition, the cause of the condition, and the route of administration.

The actual effective amount of the active compounds described hereinalso depends on the specific compound, and on the amount of fatreduction desired. The selection of the appropriate dose is well withinthe knowledge of the skilled artisan upon a reading of the presentdisclosure and based on the general knowledge of the skilled artisan.For example, in some subjects a reduction in body fat percentage to bein a range of about 25-31% from a higher percentage in females, and tobe a range of about 15-24% from a higher percentage in males, can be adesired goal. Further lowering can be desirable and can be discussedbetween the subject and their healthcare provider so as to reduce fat ina medically safe manner. The dosage amounts and treatment duration canthen be selected based on the subject's goal and the healthcareprovider's recommendation based on the medical knowledge of thehealthcare provider. As another example, the amount of fat reduction canbe an amount that results in at least about a 5% drop in body weight. Asa further example, the amount of fat reduction can be an amount toresult in a visible change in fat deposits (for example a visualreduction of submental fat, cellulite, abdominal fat, or waist fat). Thecompound can be administered in an effective amount until a desiredvisible change is achieved.

Unless indicated otherwise herein, the term “about” is intended toinclude values (e.g., weight percentages) proximate to the recited rangethat are equivalent (e.g. bioequivalent) in terms of the functionalityof the individual ingredient (e.g. active ingredient or excipient), thecomposition, or the embodiment. Furthermore, as will be understood by askilled artisan, all numbers, including those expressing quantities ofingredients, properties such as molecular weight, reaction conditions,and so forth, are approximations and are understood as being optionallymodified in all instances by the term “about.” These values can varydepending upon the desired properties sought to be obtained by thoseskilled in the art utilizing the teachings of the descriptions herein.It is also understood that such values inherently contain variabilitynecessarily resulting from the standard deviations found in theirrespective testing measurements.

Other therapeutically efficient amounts will be apparent to a skilledperson upon a reading of the present disclosure. For example, a skilledperson can determine the maximum safe dosage for healthy subjects basedon the dosages used in animal studies by routine methods (see, e.g.Dept. of Health and Human Services “Guidance For Industry: Estimatingthe Maximum Safe Starting Dose in Initial Clinical Trials forTherapeutics in Adult Healthy Volunteers”), and then administer tosubjects in need thereof various dosages below the maximum safe dosageby routine methods and experimentation until a dosage which results in adesirable effect (e.g. fat reduction without significant or additionalhair growth) is reached. Exemplary effective amounts can be about 0.05mg/kg to about 5 mg/kg. Additional effective amounts can be about 0.05mg/kg to 1 mg/kg, 0.05 mg/kg to about 2 mg/kg, about 0.05 mg/kg to about3 mg/kg, and about 0.05 mg/kg to about 4 mg/kg.

Still additional therapeutically effective amounts can be about 0.05mg/kg to about 0.15 mg/kg, about 0.15 mg/kg to about 0.27 mg/kg, about0.27 mg/kg to about 0.39 mg/kg, about 0.39 mg/kg to about 0.51 mg/kg,about 0.51 mg/kg to about 0.63 mg/kg, about 0.63 mg/kg to about 0.75mg/kg, about 0.75 mg/kg to about 0.87 mg/kg, about 0.87 mg/kg to about0.99 mg/kg, about 0.99 mg/kg to about 1.11 mg/kg, about 1.11 mg/kg toabout 1.23 mg/kg, about 1.23 mg/kg to about 1.35 mg/kg, about 1.35 mg/kgto about 1.47 mg/kg, about 1.47 mg/kg to about 1.59 mg/kg, about 1.59mg/kg to about 1.71 mg/kg, about 1.71 mg/kg to about 1.83 mg/kg, about1.83 mg/kg to about 1.95 mg/kg, about 1.95 mg/kg to about 2.07 mg/kg,about 2.07 mg/kg to about 2.19 mg/kg, about 2.19 mg/kg to about 2.31mg/kg, about 2.31 mg/kg to about 2.43 mg/kg, about 2.43 mg/kg to about2.55 mg/kg, about 2.55 mg/kg to about 2.67 mg/kg, about 2.67 mg/kg toabout 2.79 mg/kg, about 2.79 mg/kg to about 2.91 mg/kg, about 2.91 mg/kgto about 3.03 mg/kg, about 3.03 mg/kg to about 3.15 mg/kg, about 3.15mg/kg to about 3.27 mg/kg, about 3.27 mg/kg to about 3.39 mg/kg, about3.39 mg/kg to about 3.51 mg/kg, about 3.51 mg/kg to about 3.63 mg/kg,about 3.63 mg/kg to about 3.75 mg/kg, about 3.75 mg/kg to about 3.87mg/kg, about 3.87 mg/kg to about 3.99 mg/kg, about 3.99 mg/kg to about4.11 mg/kg, about 4.11 mg/kg to about 4.23 mg/kg, about 4.23 mg/kg toabout 4.35 mg/kg, about 4.35 mg/kg to about 4.47 mg/kg, about 4.47 mg/kgto about 4.59 mg/kg, about 4.59 mg/kg to about 4.71 mg/kg, about 4.71mg/kg to about 4.83 mg/kg, about 4.83 mg/kg to about 4.95 mg/kg, andabout 4.95 mg/kg to about 5 mg/kg.

The therapeutically efficient amount can be present in a formulation(e.g. for topical administration) at between about 0.01 and about 5%(w/v). In some embodiments, the therapeutically effective amount in theformulation can be from about 0.01 to about 1%, about 0.01 to about 2%,about 0.01 to about 3%, and about 0.01 to about 4%. In otherembodiments, the therapeutically effective amount in the formulation canbe from about 0.01 to about 1%, about 1 to about 2%, about 2 to about3%, about 3 to about 4%, about 4 to about 5%.

In other embodiments, the therapeutically effective amount in theformulation can be from about 0.01 to about 0.06%, about 0.06 to about0.11%, about 0.11 to about 0.16%, about 0.16 to about 0.21%, about 0.21to about 0.26%, about 0.26 to about 0.31%, about 0.31 to about 0.36%,about 0.36 to about 0.41%, about 0.41 to about 0.46%, about 0.46 toabout 0.51%, about 0.51 to about 0.56%, about 0.56 to about 0.61%, about0.61 to about 0.66%, about 0.66 to about 0.71%, about 0.71 to about0.76%, about 0.76 to about 0.81%, about 0.81 to about 0.86%, about 0.86to about 0.91%, about 0.91 to about 0.96%, about 0.96 to about 1.01%,about 1.01 to about 1.06%, about 1.06 to about 1.11%, about 1.11 toabout 1.16%, about 1.16 to about 1.21%, about 1.21 to about 1.26%, about1.26 to about 1.31%, about 1.31 to about 1.36%, about 1.36 to about1.41%, about 1.41 to about 1.46%, about 1.46 to about 1.51%, about 1.51to about 1.56%, about 1.56 to about 1.61%, about 1.61 to about 1.66%,about 1.66 to about 1.71%, about 1.71 to about 1.76%, about 1.76 toabout 1.81%, about 1.81 to about 1.86%, about 1.86 to about 1.91%, about1.91 to about 1.96%, about 1.96 to about 2.01%, about 2.01 to about2.06%, about 2.06 to about 2.11%, about 2.11 to about 2.16%, about 2.16to about 2.21%, about 2.21 to about 2.26%, about 2.26 to about 2.31%,about 2.31 to about 2.36%, about 2.36 to about 2.41%, about 2.41 toabout 2.46%, about 2.46 to about 2.51%, about 2.51 to about 2.56%, about2.56 to about 2.61%, about 2.61 to about 2.66%, about 2.66 to about2.71%, about 2.71 to about 2.76%, about 2.76 to about 2.81%, about 2.81to about 2.86%, about 2.86 to about 2.91%, about 2.91 to about 2.96%,about 2.96 to about 3.01%, about 3.01 to about 3.06%, about 3.06 toabout 3.11%, about 3.11 to about 3.16%, about 3.16 to about 3.21%, about3.21 to about 3.26%, about 3.26 to about 3.31%, about 3.31 to about3.36%, about 3.36 to about 3.41%, about 3.41 to about 3.46%, about 3.46to about 3.51%, about 3.51 to about 3.56%, about 3.56 to about 3.61%,about 3.61 to about 3.66%, about 3.66 to about 3.71%, about 3.71 toabout 3.76%, about 3.76 to about 3.81%, about 3.81 to about 3.86%, about3.86 to about 3.91%, about 3.91 to about 3.96%, about 3.96 to about4.01%, about 4.01 to about 4.06%, about 4.06 to about 4.11%, about 4.11to about 4.16%, about 4.16 to about 4.21%, about 4.21 to about 4.26%,about 4.26 to about 4.31%, about 4.31 to about 4.36%, about 4.36 toabout 4.41%, about 4.41 to about 4.46%, about 4.46 to about 4.51%, about4.51 to about 4.56%, about 4.56 to about 4.61%, about 4.61 to about4.66%, about 4.66 to about 4.71%, about 4.71 to about 4.76%, about 4.76to about 4.81%, about 4.81 to about 4.86%, about 4.86 to about 4.91%,about 4.91 to about 4.96%, and about 4.96 to about 5% (w/v).

The therapeutically effective amount can be administered according to adosing frequency that is identifiable to a skilled person during a timeperiod that is also identifiable to a skilled person. The term “dosingfrequency” as used herein, refers to the number of times the compoundsdescribed herein are administered to a subject. Exemplary dosingfrequencies include administering the effective amount at discrete timesduring a day such as, for example, once a day (QD), twice a day (BID),three times a day (TID), four times a day (QID), and others identifiableto a skilled person. Other exemplary dosing frequencies includecontinuous dosing, for example by intravenous infusion, use of a drugpump, use of a transdermal patch, or other methods of continuous dosingidentifiable to a skilled person.

The therapeutically effective amount can be administered at a desireddosing frequency for a time period identifiable to a skilled person. Forexample, a therapeutically effective can be administered once or twice aday (or at another dosing frequency identifiable to a skilled person)for a set period of time (e.g. seven to fourteen days, two to fourweeks, one to six months, or for another time period identifiable to askilled person). As another example, a therapeutically effective amountcan be administered once or twice a day (or at another dosing frequencyidentifiable to a skilled person) for a non-predetermined period oftime. A skilled person can determine at various points during the periodof time if the administration of the effective amount is to be continued(e.g., if a desired outcome such as a particular amount of fat loss hasbeen achieved and administration of the effective amount is not requiredand/or desired anymore).

Some compounds described herein have at least one asymmetric center intheir structure. This asymmetric center can be present in an R or Sconfiguration, said R and S notation is used in correspondence with therules described in Pure Applied Chem. 1976, 45, 11-13.

Reference to a compound or compounds described herein is intended toencompass that compound in each of its possible isomeric forms andmixtures thereof unless the particular isomeric form is referred tospecifically.

Synthesis of compounds described herein (e.g. Formulas I and II) can beachieved by known methods, for example, those recited in WO1996/036599and WO1997/031895.

In some embodiments, the compounds described herein can be used in themanufacture of a medicament for fat reduction in a subject.

Another embodiment is a kit comprising a composition comprising acompound disclosed herein, a container, and instructions foradministration of said composition to a subject for fat reduction.

EXAMPLES

The following examples are intended only to illustrate the presentdisclosure and should in no way be construed as limiting the presentdisclosure.

Example 1: Lack of Activity on Human Dermal Papilla Cells

In this example, the lack of activity of the compound according toFormulas I and II is shown. In general, bimatoprost or the compound ofFormula I was incubated at concentrations from 10⁻⁵ and 10⁻¹² M withhuman dermal papilla cells in an in vitro impedance assay. Bimatoprostcaused a down-ward signal (relaxation) or reduction in impedance, whilethe compound of Formula I did not produce any detectable effect. Threeindependent experiments were performed.

More specifically, Human Hair Dermal Papilla Cells (HHDPCs) wereexpanded in specific culture medium (Promo Cell). After growing, thecells were seeded on 96 well plates coated with poly-L-Lysine at 20,000cells per well. Cells were incubated during 24 hours at 37° C., 5% CO₂.The day of the experiment, the culture medium was eliminated andreplaced by HBSS containing BSA, 0.1% v/v. The cells were incubated inthe CellKey system, for 4 hours at 28° C. before using to testcompounds. Bimatoprost, the compound of Formula I, and/or the compoundof Formula II were tested at 7 concentrations from 10⁻⁵ to 10⁻¹² M. Thechange in impedance was measured over 20 minutes. The background wassubtracted and the impedance signal was normalized to the reference(bimatoprost) for the compound of Formula I and the compound of FormulaII. The data presented is in triplicate and of three independentexperiments.

The results of the assay can be seen in FIGS. 1A-1D. Bimatoprost showsactivity on primary human dermal papilla cells whereas compounds ofFormulas I and II do not show activity (see FIGS. 2A-2D for the compoundof Formula II and FIGS. 3A-3D for the compound of Formula I). Theseresults indicate that the compounds according to Formulas I and II donot exhibit activity and thus do not activate receptors present on thesecells. These cells are the primary driver of hair growth, for whichstimulation by bimatoprost and other compounds, such as minoxidil,mediate their action (see, e.g., Khidhir et al. FASB J. 2013, 27,557-567; Messenger et al. Br. J. Dermatol. 2004, 150, 186-194; and Li etal. J. Invest. Dermatol. 2001, 117, 1594-1600) the hair-growthassociated with molecules such as bimatoprost.

Example 2: Lack of Acceleration of Time to Full Hair Growth

In this Example, it is shown that the compound of Formula I does notaccelerate the time to full hair growth. In general, the compound offormula I and bimatoprost were prepared in the minoxidil formulation(50% Propylene Glycol; 30% Ethyl Alcohol) at a concentration of 0.03%for both compounds. Both compounds were given topically to shaved backsof mice (as described in Khidhir et al. FASEB J. 2013 27(2):557-567).

More specifically, female C57BL/6J mice (cat. no. 000664; JacksonLaboratory, Bar Harbor, Me., USA), aged 7 weeks, were randomlydistributed into 4 groups to avoid any sibling bias and housed in groupsof 5 with standard diet food pellets and water available ad libitum.Initially, dorsal hairs were removed externally by shaving (˜2×6 cm)using an electric trimmer (Wahl Stylique Designer/Liner pet trimmer;919179; Petco, San Diego, Calif., USA) revealing pink skin. From thenext day, termed d 0 (zero), each mouse was treated topically witheither the vehicle alone (ethanol:propylene glycol:water 3:5:2) or 0.03,0.10, or 0.30% bimatoprost in the vehicle for d 14; 10 mice were usedfor each condition. The appropriate solution (70 μl) was rubbed gentlyinto the dorsal skin of each mouse daily; new gloves were worn for eachtreatment type. Hair growth was recorded daily for each animal for d 42,and dorsal photographs were taken at d 0, 7, 14, 17, 21, 24, 28, 31, 35,and 42. The first day of anagen, defined as the first day when visibledarkness could be seen that subsequently increased and progressed tovisible black hair, was recorded for each animal. To calculate meanvalues, the first day of growth was designated as d 43 for any mouseshowing no hair growth signs by the last day. The day when the shaveddorsal area was fully covered with new black hairs, i.e., there was nopink skin remaining and no visible difference in hair length to theadjacent unshaved areas, was also recorded. This assay has beensuccessfully used to evaluate compounds that cause hair growth (see,e.g., Khidhir et al. FASB J. 2013, 27, 557-567; Plikus et al. J. Invest.Dermatol. 2008, 128, 1071-1080).

The results of this example can be seen FIGS. 4A and 4B. While Formula Iappears to boost the start to hair re-growth it does not result in fullhair re-growth. Administration of bimatoprost accelerates both time tostart and full hair re-growth. This indicates that the compound ofFormula I does not re-grow hair in this mouse model.

Example 3: Activity on Pre-Adipocytes

In this example it is shown that the compounds of Formulas I and IIstimulates pre-adipocytes. In general, the compounds of Formulas I andII and bimatoprost were tested in the xCelligence impedance assay. Bothbimatoprost and the compounds of Formulas I and II stimulate prostamidereceptors on human pre-adipocytes.

More specifically, pre-adipocytes were purchased from Lonza. Cells wereexpanded in Pre-adipocyte maintenance medium (Lonza, Switzerland). Onceexpanded, 10,000 cells per well were plated into xCelligence 96-wellplates and incubated at 37° C. for 72 hours until the impedance signalnormalized. Once normalized, media was switched to serum-freepre-adipocyte media (PDM-2) for 2 hours at 37° C. Once the signalstabilized for 30 minutes, compounds were added at 7 concentrations(10⁻⁵ to 10⁻⁹ M) and the signal measured for 20 minutes. The change inimpedance was normalized to the background reading (buffer alone) andthe EC₅₀ calculations were determined using prism software.

TABLE 1 Compound xCelligence EC₅₀ Bimatoprost 7.84 × 10⁻⁷ Formula I 1.07× 10⁻⁸ Formula II 1.17 × 10⁻⁸

The results in Table 1 show that formula I and II are relatively morepotent than bimatoprost on human pre-adipocytes.

In addition, 3T3-L1 mouse pre-adipocytes were induced to differentiatein the presence of bimatoprost, compounds of formula I and II for 3 daysat the concentrations indicated. After 3 days the media and compoundswere removed and replaced with adipocyte maintenance media. The mediawas change every 2 days following up to 8 days after initial induction.Lipid content was measured by using adipored dye (Lonza) and measured byfluorescent plate reader. The differentiation index was calculated as aratio of differentiated wells to undifferentiated controls. 100% isdefined as the differentiation index of differentiated control wellscontaining vehicle only. Differentiation media conditions were inDMEM/F-12 media containing insulin, isobutylmethylxanthine, anddexamethasone. Maintenance media contained insulin and dexamethasoneonly. The results are shown in FIG. 6.

Example 4: Reduction of Weight Gain and Lowering of Mean Body Weights

In this example it is shown that the compound of Formula I whenadministration orally daily results in reduction in weight gain andlower mean body weights. In general males rats (n=17-20) and female rats(n=17-20) were administered orally (by gavage) daily for 6 months at thedoses indicated below. The compound of Formula I inhibited weight gainin both male and females at 5 mg/kg/day, and 0.3 mg/kg/day for females(p<0.05). The effect was somewhat reversible after administration ofdrug ceased.

More specifically, male and female rats (Hsd: Sprague Dawley) wereassigned to four groups and administered test article formulations(0.03, 0.3, or 5 mg/kg/day compound of Formula I) or vehicle controlarticle (0 mg/mL [0%] compound of Formula I [placebo] containing 3%hydroxypropyl-β-cyclodextrin [30 mg/mL (150 mg/kg/day)] and 10 mMphosphate buffered saline) via oral gavage once daily for 26 weeks at avolume of 5 mL/kg. Animals were sacrificed at the end of the dosingphase (up to 15 animals/sex/group) or at the end of the 4-week recoveryphase (5 animals/sex/group in dose groups given vehicle control articleand 5 mg/kg/day the compound of Formula I). These animals were on arestricted diet due to the length of the study. Weights were measuredonce per week.

The results of the example are shown in FIGS. 5A (male rats) and 5B(female rats). As can be seen from the graphs, administration of FormulaI daily resulted in a decrease in body weight gain starting at 14 days.This persistence in reduction continued throughout the treatment period,up to 183 days. Following the washout period, animals recovered aportion of the weight deficit they exhibited during the treatmentperiod. This data indicates the weight loss is reversible. A skilledperson will understand upon a reading of the present disclosure thatdosages can be from about 0.05 mg/kg to about 5 mg/kg, and also fromabout 0.3 mg/kg to about 5 mg/kg.

Throughout this specification reference is made to publications such asUS and foreign patent applications, journal articles, book chapters, andothers. All such publications are expressly incorporated by reference intheir entirety, including supplemental/supporting information sectionspublished with the corresponding references, for all purposes unlessotherwise indicated. To the extent that any recitations in theincorporated references conflict with any recitations herein, therecitations herein will control.

The foregoing descriptions details specific methods that can be employedto reduce fat, and in particular reduce fat without significant hairgrowth and/or additional hair growth, and represents the best modecontemplated. It should not be construed as limiting the overall scopehereof; rather, the ambit of the present disclosure is to be governedonly by the lawful construction of the appended claims.

What is claimed is:
 1. A method of reducing body fat in a subjectcomprising administering topically to a subject in need thereof aneffective amount compound of Formula I, a compound of Formula II, or amixture thereof:


2. The method of claim 1, wherein an effective amount of the compound offormula I is administered.
 3. The method of claim 1, wherein aneffective amount of the compound of formula II is administered.
 4. Themethod of claim 1, wherein the compound or mixture of compounds isadministered in an amount effective for reducing fat without causingsubstantial hair growth at the site of fat reduction.
 5. The method ofclaim 1, wherein the compound or mixture of compounds is administered inan amount effective for reducing fat without causing additional hairgrowth at the site of fat reduction.
 6. The method of claim 1, whereinthe compound or mixture of compounds is administered in an amount ofabout 0.05 mg/kg of body weight to about 5 mg/kg of body weight.
 7. Themethod of claim 1, wherein the compound or mixture of compounds isadministered in an amount of about 0.3 mg/kg of body weight to about 5mg/kg of body weight.
 8. The method of claim 1, wherein the compound ormixture of compounds is administered to at least one of the subject'ssubmental region, thighs, abdomen, or waist.
 9. The method of claim 1,wherein the compound or mixture of compounds is administered locally toa fat deposit.